Bottom Line:
The effect of stress on PER1 and FOS was modulated by time of day and, in the case of daily restraint, by predictability.Furthermore, the results show that the mechanisms that control PER1 and FOS expression in CEAl and BNSTov are uniquely sensitive to differences in the type of stressor.Finally, the finding that the effect of stress on PER1 parallels its effect on FOS supports the idea that Per1 functions as an immediate-early gene.

ABSTRACTStressful events can disrupt circadian rhythms in mammals but mechanisms underlying this disruption remain largely unknown. One hypothesis is that stress alters circadian protein expression in the forebrain, leading to functional dysregulation of the brain circadian network and consequent disruption of circadian physiological and behavioral rhythms. Here we characterized the effects of several different stressors on the expression of the core clock protein, PER1 and the activity marker, FOS in select forebrain and hypothalamic nuclei in rats. We found that acute exposure to processive stressors, restraint and forced swim, elevated PER1 and FOS expression in the paraventricular and dorsomedial hypothalamic nuclei and piriform cortex but suppressed PER1 and FOS levels exclusively in the central nucleus of the amygdala (CEAl) and oval nucleus of the bed nucleus of the stria terminalis (BNSTov). Conversely, systemic stressors, interleukin-1β and 2-Deoxy-D-glucose, increased PER1 and FOS levels in all regions studied, including the CEAl and BNSTov. PER1 levels in the suprachiasmatic nucleus (SCN), the master pacemaker, were unaffected by any of the stress manipulations. The effect of stress on PER1 and FOS was modulated by time of day and, in the case of daily restraint, by predictability. These results demonstrate that the expression of PER1 in the forebrain is modulated by stress, consistent with the hypothesis that PER1 serves as a link between stress and the brain circadian network. Furthermore, the results show that the mechanisms that control PER1 and FOS expression in CEAl and BNSTov are uniquely sensitive to differences in the type of stressor. Finally, the finding that the effect of stress on PER1 parallels its effect on FOS supports the idea that Per1 functions as an immediate-early gene. Our observations point to a novel role for PER1 as a key player in the interface between stress and circadian rhythms.

pone-0111166-g009: The effect of acute nighttime stress on PER1 and FOS expression.Rats were exposed to 30 min restraint or treatment with 250 mg/kg 2DG, administered during the nighttime at ZT14, and then killed at ZT15. Means ± SEM of immunoreactive (IR) cells are shown, n = 4 per group; * significant difference from corresponding control group, p<0.05.

Mentions:
Plasma CORT varied across all 3 groups (F2,9 = 7.68, p<0.05; Fig. 8). CORT levels were significantly higher than control levels following the 2DG challenge (Fig. 8, p<0.05), though not after restraint. Neither nighttime restraint nor 2DG treatment had any effect on PER1 or FOS levels in the SCN (Fig. 9A). Similarly, there were no effects of either stressor on PER1 expression in the PVN. FOS levels in the PVN were significantly increased only in response to restraint (F2,8 = 8.11, p<0.01; Fig. 9B). In the DMH (Fig. 9C) and in the Pi (Fig. 9D), PER1 and FOS expression were significantly increased by both stressors (DMH: PER1, F2,9 = 7.38, p<0.05; FOS, F2,9 = 9.13, p<0.01; Pi: PER1, F2,9 = 25.93, p<0.01; FOS, F2,9 = 65.44, p<0.0001). Nighttime restraint, though not 2DG, significantly suppressed PER1 levels in the CEAl (F2,9 = 7.05, p<0.01; Fig. 9F). There was no effect of either stressor on PER1 expression in the BNSTov (Fig. 9E). FOS expression varied significantly across the stress groups in the BNSTov (F2,9 = 21.07, p<0.01, Fig. 9E) and CEAl (F2,9 = 51.57, p<0.01; Fig. 9F). Both regions exhibited an increase in FOS expression following exposure to 2DG but not restraint (Figs. 9E, F).

pone-0111166-g009: The effect of acute nighttime stress on PER1 and FOS expression.Rats were exposed to 30 min restraint or treatment with 250 mg/kg 2DG, administered during the nighttime at ZT14, and then killed at ZT15. Means ± SEM of immunoreactive (IR) cells are shown, n = 4 per group; * significant difference from corresponding control group, p<0.05.

Mentions:
Plasma CORT varied across all 3 groups (F2,9 = 7.68, p<0.05; Fig. 8). CORT levels were significantly higher than control levels following the 2DG challenge (Fig. 8, p<0.05), though not after restraint. Neither nighttime restraint nor 2DG treatment had any effect on PER1 or FOS levels in the SCN (Fig. 9A). Similarly, there were no effects of either stressor on PER1 expression in the PVN. FOS levels in the PVN were significantly increased only in response to restraint (F2,8 = 8.11, p<0.01; Fig. 9B). In the DMH (Fig. 9C) and in the Pi (Fig. 9D), PER1 and FOS expression were significantly increased by both stressors (DMH: PER1, F2,9 = 7.38, p<0.05; FOS, F2,9 = 9.13, p<0.01; Pi: PER1, F2,9 = 25.93, p<0.01; FOS, F2,9 = 65.44, p<0.0001). Nighttime restraint, though not 2DG, significantly suppressed PER1 levels in the CEAl (F2,9 = 7.05, p<0.01; Fig. 9F). There was no effect of either stressor on PER1 expression in the BNSTov (Fig. 9E). FOS expression varied significantly across the stress groups in the BNSTov (F2,9 = 21.07, p<0.01, Fig. 9E) and CEAl (F2,9 = 51.57, p<0.01; Fig. 9F). Both regions exhibited an increase in FOS expression following exposure to 2DG but not restraint (Figs. 9E, F).

Bottom Line:
The effect of stress on PER1 and FOS was modulated by time of day and, in the case of daily restraint, by predictability.Furthermore, the results show that the mechanisms that control PER1 and FOS expression in CEAl and BNSTov are uniquely sensitive to differences in the type of stressor.Finally, the finding that the effect of stress on PER1 parallels its effect on FOS supports the idea that Per1 functions as an immediate-early gene.

ABSTRACTStressful events can disrupt circadian rhythms in mammals but mechanisms underlying this disruption remain largely unknown. One hypothesis is that stress alters circadian protein expression in the forebrain, leading to functional dysregulation of the brain circadian network and consequent disruption of circadian physiological and behavioral rhythms. Here we characterized the effects of several different stressors on the expression of the core clock protein, PER1 and the activity marker, FOS in select forebrain and hypothalamic nuclei in rats. We found that acute exposure to processive stressors, restraint and forced swim, elevated PER1 and FOS expression in the paraventricular and dorsomedial hypothalamic nuclei and piriform cortex but suppressed PER1 and FOS levels exclusively in the central nucleus of the amygdala (CEAl) and oval nucleus of the bed nucleus of the stria terminalis (BNSTov). Conversely, systemic stressors, interleukin-1β and 2-Deoxy-D-glucose, increased PER1 and FOS levels in all regions studied, including the CEAl and BNSTov. PER1 levels in the suprachiasmatic nucleus (SCN), the master pacemaker, were unaffected by any of the stress manipulations. The effect of stress on PER1 and FOS was modulated by time of day and, in the case of daily restraint, by predictability. These results demonstrate that the expression of PER1 in the forebrain is modulated by stress, consistent with the hypothesis that PER1 serves as a link between stress and the brain circadian network. Furthermore, the results show that the mechanisms that control PER1 and FOS expression in CEAl and BNSTov are uniquely sensitive to differences in the type of stressor. Finally, the finding that the effect of stress on PER1 parallels its effect on FOS supports the idea that Per1 functions as an immediate-early gene. Our observations point to a novel role for PER1 as a key player in the interface between stress and circadian rhythms.